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971.
Shaopeng Chen Junkang Qiu Chuan Chen Chunchun Liu Yuheng Liu Lili An Junying Jia Jie Tang Lijun Wu Haiying Hang 《蛋白质与细胞》2012,3(6):460
Activation-induced cytidine deaminase (AID) is required for the generation of antibody diversity through initiating both somatic hypermutation (SHM) and class switch recombination. A few research groups have successfully used the feature of AID for generating mutant libraries in directed evolution of target proteins in B cells in vitro . B cells, cultured in suspension, are not convenient for transfection and cloning. In this study, we established an AID-based mutant accumulation and sorting system in adherent human cells. Mouse AID gene was first transfected into the human non-small cell lung carcinoma H1299 cells, and a stable cell clone (H1299-AID) was selected. Afterwards, anti-hTNF-αscFv (ATscFv ) was transfected into H1299-AID cells and ATscFv was displayed on the surface of H1299-AID cells. By 4-round amplification/flow cytometric sorting for cells with the highest affinities to hTNF-alpha, two ATscFv mutant gene clones were isolated. Compared with the wild type ATscFv, the two mutants were much more efficient in neutralizing cytotoxicity of hTNF-alpha. The results indicate that directed evolution by somatic hypermutation can be carried out in adherent non-B cells, which makes directed evolution in mammalian cells easier and more efficient. 相似文献
972.
973.
Jian-Ping An Xin Liu Lai-Qing Song Chun-Xiang You Xiao-Fei Wang Yu-Jin Hao 《Journal of Plant Biology》2017,60(2):137-145
RING-finger-containing E3 ubiquitin ligases play important roles in plant response to biotic and abiotoc stresses. In this study, through homology analysis, a Malus× domestica MYB30-Interacting E3 Ligase 1 gene, MdMIEL1, was identified and subsequently cloned from apple ‘Gala’ (Malus×domestica). MdMIEL1 contained a zinc finger domain close to N-terminus and a RING finger domain close to Cterminus. Expression of MdMIEL1 was significantly induced by NaCl and H2O2 treatments. Further study demonstrated that the MdMIEL1-overexpressing Arabidopsis and apple calli were less tolerance to salt stress than wild-type control. In addition, transgenic plants had higher levels of reactive oxygen species (ROS) (H2O2 and O2 –). And transgenic Arabidopsis and apple calli exhibited more sensitive phenotype to H2O2 treatment, which was associated with increased levels of ROS. These findings indicate MdMIEL1 is an important regulator involved in plant response to salt and oxidative stresses tolerance. 相似文献
974.
Jung Hye Hwang Sang Mi An Seul Gi Kwon Da Hye Park Tae Wan Kim Deok Gyung Kang 《Animal biotechnology》2017,28(4):288-293
Postmortem pH is a main factor influencing the meat quality in pigs. This study investigated the association of postmortem pH with single-nucleotide polymorphisms (SNPs) in the fourth member of the short-chain dehydrogenase/reductase family (DHRS4), the first member of serpin peptidase inhibitor, clade G (complement inhibitor) (SERPING1), and the apolipoprotein R precursor (APOR) genes in Berkshire pigs. The study included 437 pigs, and genotyping was conducted using the GoldenGate Assay (Illumina, San Diego, CA, USA). DHRS4, SERPING1, and APOR polymorphisms were significantly associated with pH45 or pH24 (p?0.05). SERPING1 was also statistically significantly associated with water holding capacity (p?0.05), which is closely associated with postmortem pH. These results suggest that SNPs in the DHRS4, SERPING1, and APOR genes have potential for use as genetic markers for the meat quality in pigs. 相似文献
975.
Tongtong Liu Yuling An Kai Liu Fangfang Wang Chupeng Xie Ying Zhang Xin Guan Jichun Tian Jiansheng Chen 《Molecular breeding : new strategies in plant improvement》2017,37(3):41
Dissecting the genetic basis for the traits of northern-style Chinese steamed bread (NCSB) is of great significance for wheat quality breeding. Quantitative trait loci (QTLs) for the processing quality of NCSB were studied using a recombinant inbred line (RIL) consisting of 173 lines derived from a “Shannong01–35 × Gaocheng9411” cross. Twenty-four putative additive QTLs were detected on chromosomes 1A, 1B, 1D, 3A, 3B, 4A, 4B, 5B, 6B, and 7B. Of these QTLs, QTex1A.1-27, QHei5B.5-488, and QGum4B.4-17 had the highest contribution and accounted for 9.33, 10.9, and 12.0% of the phenotypic variations, respectively. Several co-located QTLs with additive effects were detected on chromosomes 1A, 1D, 4B, and 5B. Two clusters (RFL_CONTIG2160_524-WSNP_CAP12_C2438_1180601 and EX_C101685_705-RAC875_C27536_611) for height, total score, and texture and for chewiness, gumminess, and hardness were detected on chromosomes 1A and 4B, respectively. Two QTLs for chewiness and hardness (QCh1D-4, QHa1D-4) with additive effects were detected; these alleles could be good targets for improving the processing quality of steamed bread from common wheat (Triticum aestivum L.). In addition, QTLs for wheat flour quality and the associated correlations with NCSB were simultaneously analyzed. Negative correlations were detected between chewiness and the wet/dry gluten content (WGC/DGC) or protein content. Two QTLs (QCh4B.4-17 and QPr4B.4-17) and three QTLs (QCh4B.4-13, QWG4B.4-13, and QDG4B.4-13) clustered in the same chromosomal region. The detected QTL clusters should be further investigated during wheat breeding and could be used by breeders to improve wheat quality and especially the processing quality of NCSB. 相似文献
976.
Li Lulu An Meiling Qu Changfeng Zheng Zhou Wang Yibin Liu Fangming He Yingying He Xiaodong Miao Jinlai 《Extremophiles : life under extreme conditions》2017,21(4):817-827
Extremophiles - Major intrinsic proteins (MIPs) form channels facilitating the passive transport of water and other small polar molecules across membranes. In this study, the complete open reading... 相似文献
977.
S,N co‐doped carbon quantum dots (N,S‐CQDs) with super high quantum yield (79%) were prepared by the hydrothermal method and characterized by transmission electron microscopy, photoluminescence, UV–Vis spectroscopy and Fourier transformed infrared spectroscopy. N,S‐CQDs can enhance the chemiluminescence intensity of a luminol–H2O2 system. The possible mechanism of the luminol–H2O2–(N,S‐CQDs) was illustrated by using chemiluminescence, photoluminescence and ultraviolet analysis. Ranitidine can quench the chemiluminescence intensity of a luminol–H2O2–N,S‐CQDs system. So, a novel flow‐injection chemiluminescence method was designed to determine ranitidine within a linear range of 0.5–50 μg ml?1 and a detection limit of 0.12 μg ml?1. The method shows promising application prospects. 相似文献
978.
Two simple colorimetric receptors PS and PP (thiophene and pyridine appended derivative) were prepared and their cation sensing properties were investigated. Receptors PS and PP displayed a selective colorimetric change (from colorless to orange) upon binding to Fe3+ in MeOH solution. The association constants for receptors PS–F e3+ and PP–F e3+ in MeOH were determined to be 1.15 × 106 and 4.31 × 106 M?1, respectively, using Hill plots. The detection limits of PS and PP were 490.7 ppm and 393.7 ppm, respectively. 相似文献
979.
Quoc Tuc Dinh Truong An Nguyen Elodie Moreau-Guigon Fabrice Alliot Marie-Jeanne Teil Martine Blanchard 《Soil & Sediment Contamination》2017,26(3):247-258
This work presents the optimization of analytical procedures for the determination of two antibiotics, oxolinic acid (OA) and flumequine (FL), in bed sediment, river water, and soil samples. Three extraction methods (microwave-assisted extraction (MAE), ultrasonication, and reflux) were tested, and the highest recoveries were obtained with MAE (94 ± 3% and 95 ± 3% for OA and FL, respectively). A solid-phase extraction (SPE) clean-up step was optimized by comparing two polymeric sorbents: Oasis HLB and Oasis MAX. The final extracts were analyzed by liquid chromatography with fluorimetric detection. Limits of detection (LOD) obtained for OA and FL in soil and sediment ranged from 0.3 to 0.5 µg kg?1. Meanwhile, a novel SPE procedure was also implemented for OA and FL determination in river water samples. It also relied on the use of Oasis MAX, and recovery rates were in the range 90–94%; LODs were 2 ng L?1 for both OA and FL. These methods were applied for the analysis of samples taken in the Seine River basin (France). The obtained results demonstrated the widespread occurrence of OA and FL, at ng L?1 and µg kg?1 levels in water and sediment/soil, respectively, and their persistence in the environment. 相似文献
980.
Traditional neonicotinoid insecticides are used worldwide. Paichongding (IPP), as a novel neonicotinoid pesticide, has been widely used in China. However, the ecotoxicity of IPP to non-target invertebrates in soil ecosystem has not been reported yet. In this study, acute toxicity of IPP to earthworm Eisenia fetida, as well as the antioxidant response after IPP exposure, was evaluated. In the filter paper contact test, the LC50 at 24 hr and 48 hr for IPP were 14.98 μg/cm2 and 7.59 μg/cm2, respectively. In artificial soil test, the LC50 (lethal concentration) at 14 days and 28 days for IPP were 541.07 mg/kg and 238.51 mg/kg, respectively. The LC50 of IPP is much higher than that of traditional neonicotinoid insecticides. However, earthworm body weight assessment demonstrated that the growth of earthworm was inhibited by extended exposure to IPP at sublethal doses. The activities of antioxidative enzymes superoxide dismutase and catalase in earthworms were significantly induced after IPP exposure. Malondialdehyde, a biomarker of lipid peroxidation, was also increased after IPP exposure. Although the results indicated that IPP had potentially adverse effect on earthworms, its toxicity was much lower than traditional neonicotinoids. 相似文献